ArticleApplied and environmental microbiology1997
An assay combining cell culture with reverse transcriptase PCR to detect and determine the infectivity of waterborne Cryptosporidium parvum.
Article in Applied and environmental microbiology, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 153 citations in OpenAlex.
- Waterborne pathogens: detection methods and challenges.Pathogens (Basel, Switzerland) · 2015Review
- Comparison of assays for sensitive and reproducible detection of cell culture-infectious Cryptosporidium parvum and Cryptosporidium hominis in drinking water.Applied and environmental microbiology · 2012Article
- Detection of viable Cryptosporidium parvum in soil by reverse transcription-real-time PCR targeting hsp70 mRNA.Applied and environmental microbiology · 2011Article
- Determining UV inactivation of Toxoplasma gondii oocysts by using cell culture and a mouse bioassay.Applied and environmental microbiology · 2010Article
- Comparative efficacy of conventional primer sets in detection of Cryptosporidium parvum for diagnostic use.Parasitology research · 2010Article
- CP2 gene as a useful viability marker for Cryptosporidium parvum.Parasitology research · 2008Article
- Aged HCT-8 cell monolayers support Cryptosporidium parvum infection.Applied and environmental microbiology · 2007Article
- Evaluation of two DNA template preparation methods for post-immunomagnetic separation detection of Cryptosporidium parvum in foods and beverages by PCR.Applied and environmental microbiology · 2007Article
- An evaluation of primers amplifying DNA targets for the detection of Cryptosporidium spp. using C. parvum HNJ-1 Japanese isolate in water samples.Parasitology research · 2007Article
- Quantitative-PCR assessment of Cryptosporidium parvum cell culture infection.Applied and environmental microbiology · 2005Article
- Effects of the Norwegian winter environment on Giardia cysts and Cryptosporidium oocysts.Microbial ecology · 2004Article
- Molecular characterization of isolates of waterborne Cryptosporidium spp. collected during an outbreak of gastroenteritis in South Burgundy, France.Journal of clinical microbiology · 2003Article
- Giardia cysts in wastewater treatment plants in Italy.Applied and environmental microbiology · 2003Article
- Cell culture-Taqman PCR assay for evaluation of Cryptosporidium parvum disinfection.Applied and environmental microbiology · 2003Article
- Comparison of method 1623 and cell culture-PCR for detection of Cryptosporidium spp. in source waters.Applied and environmental microbiology · 2003Article
- Real-time PCR method for detection of Encephalitozoon intestinalis from stool specimens.Journal of clinical microbiology · 2002Article
- Comparison of in vitro cell culture and a mouse assay for measuring infectivity of Cryptosporidium parvum.Applied and environmental microbiology · 2002Article
- In vitro cultivation of cryptosporidium species.Clinical microbiology reviews · 2002Review
- U.S. drinking water challenges in the twenty-first century.Environmental health perspectives · 2002Review
- Determination of pyrimidine dimers in Escherichia coli and Cryptosporidium parvum during UV light inactivation, photoreactivation, and dark repair.Applied and environmental microbiology · 2001Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The presence of Cryptosporidium in drinking water supplies is a significant problem faced by the water industry. Although a variety of methods exist for the detection of waterborne oocysts, water utilities currently have no way of assessing the infectivity of detected oocysts and consequently are unable to accurately determine the risks posed to public health by waterborne Cryptosporidium. In this paper, the development of an infectivity assay for waterborne Cryptosporidium parvum is described. Oocysts were inoculated onto monolayers of Caco-2 cells and grown on microscope slides, and infections were detected by C. parvum specific reverse transcriptase PCR of extracted mRNA, targeting the heat shock protein 70 (hsp70) gene. A single infectious oocyst was detected by this experimental procedure. The use of concentrated samples obtained from 250 liters of finished water had no observable effect on the integrity of cell monolayers or on the infectivity of oocysts seeded into the concentrate. Intracellular developmental stages of the parasite were also detected by using fluorescently labeled antibodies. One pair of PCR primers targeting the hsp70 gene was specific for C. parvum, while a second pair recognized all species of Cryptosporidium tested. The C. parvum-specific primers amplified DNA from 1 to 10 oocysts used to seed 65 to 100 liters of concentrated environmental water samples and were compatible with multiplex PCR for the simultaneous detection of C. parvum and Giardia lambia. This paper confirms the utility of PCR for the detection of waterborne C. parvum and, most importantly, demonstrates the potential of an in vitro infectivity assay.
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